化学伝達物質としてのウシラクトフェリンの機能とその構造ユニットの解明

阐明牛乳铁蛋白作为化学信使的功能及其结构单元

• 批准号: 03660289
• 负责人: SHIMAZAKI Kei-ichi
• 金额: $ 1.41万
• 依托单位: Obihiro University of Agriculture and Veterinary Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03660289/
• 关键词: lactoferrin; milk protein; monocyte; Trypanosoma; fragments; Trypanosoma cruzi; レセプタ-

The C terminal half molecule (C lobe) of bovine lactoferrin was isolated by mild tryptic hydrolysis of lactoferrin followed by gel filtration and ion-exchange chromatography. The idntity of the fragment was established by determining its N terminal and C terminal amino acid sequences and comparing them with the amino acid sequence of intact lactoferrin.The binding properties of intact lactoferrin and its fragments have been tested with bovine monocyte and with Trypanosoma cruzi. The specific binding with bovine lactoferrin was studied. Lactoferrin isolated from mature bovine milk by chromatographic method was used and labeled by ^<125>I-Bolton-Hunter reagent. From the binding assay, it was found that bovine milk lactoferrin could bind to bovine monocyte, specifically. And intact lactoferrin showed the binding ability to Trypanosoma cruzi (amastigote form). However, C terminal half molecule of bovine lactoferrin loses the binding capacity against Trypanosoma cruzi. Amastigote of Trypanosoma cruzi was obtained from mouse embryo fibroblast culture. The binding was studied by the method of the direct immunofluorescence test using FITC-labeled anti-bovine lactoferrin antibody (rabbit).

以牛乳铁蛋白为原料，通过温和的胰酶水解、凝胶过滤和离子交换层析分离得到牛乳铁蛋白的C末端半分子(C瓣)。通过测定其N端和C端的氨基酸序列并与完整乳铁蛋白的氨基酸序列进行比较，确定了该片段的同源性。用牛单核细胞和克氏锥虫检测了完整乳铁蛋白与其片段的结合特性。研究了乳铁蛋白与牛乳铁蛋白的特异性结合。用层析法从成熟牛乳中分离出乳铁蛋白，并用~(125)&gt；i-Bolton-Hunter试剂标记。结合实验表明，牛乳乳铁蛋白能与牛单核细胞特异结合。完整的乳铁蛋白对克氏锥虫(无鞭毛体)具有结合能力。然而，牛乳铁蛋白的C端半分子失去了与克氏锥虫的结合能力。克氏锥虫无鞭毛体是从小鼠胚胎成纤维细胞培养中获得的。用FITC标记的抗牛乳铁蛋白抗体(兔)直接免疫荧光法研究其结合情况。

项目成果

K.Shimazaki and I.Kiyosawa: "Lactoferrin - Structure and Function" Bioscience and Industry. 51(1). 25-27 (1993)

K.Shimazaki 和 I.Kiyosawa：“乳铁蛋白 - 结构和功能”生物科学与工业。

島崎 敬一ほか: "Bovine monocyte separation and its interaction with lactoferrin: A preliminary study"

Keiichi Shimazaki 等人：“牛单核细胞分离及其与乳铁蛋白的相互作用：初步研究”

島崎 敬一: "Separation and Characterization of the C-terminal Half Molecule of Bovine Lactoferrin" J.Dairy Sci.76. (1993)

Keiichi Shimazaki：“牛乳铁蛋白 C 末端半分子的分离和表征”J.Dairy Sci.76（1993）。

K.Shimazakiら10名: "Separation and characterization of the C-terminal-halfmolecule of bovine lactoferrin." J.Dairy Sci.76. 946-955 (1993)

K. Shimazaki 等 10 人：“牛乳铁蛋白 C 末端半分子的分离和表征”，J.Dairy Sci.76（1993）。

K.Shimazaki et al.: "Separation and characterization of C-terminal molecule of bovine lactoferrin" J.Dairy Sci.76(8). 946-955 (1993)

K.Shimazaki 等人：“牛乳铁蛋白 C 末端分子的分离和表征”J.Dairy Sci.76(8)。